The MCIC microscopy laboratory is fully equipped to conduct biological research at the subcellular and ultrastuctural level. We house light and confocal microscopes, two transmission electron microscopes and two scanning electron microscopes. We offer a variety of sample preparation techniques and work closely with investigators to optimize the various experimental procedures. The MCIC personnel provides assistance and/or individual training in the use of the light and electron microscopes, and technical support for electron microscopy sample preparation. In collaborative projects, the MCIC staff helps design light/EM microscopy experiments and assists with the interpretation of the data. The MCIC staff also trains investigators who wish to learn microscopy techniques and conduct their own work (sample preparation and embedding for light or electron microscopy, thin and ultra-thin sectioning, etc.).
To use microscopy services, you need first to contact the MCIC staff to be trained on the equipment and/or to discuss the project. Microscope users must be trained by the facility personnel before getting access to the microscopes. You will also need to fill in, and submit a "Microscopy Work Form".
Instrumentation. Several dissecting and light microscopes available to users. The epifluorescence inverted Leica DM IRB microscope is equiped withQ Imaging Retiga 2000 cooled digital camera. The microscope has the following filters: (1) UV (Chroma A: BP 340-380/400/LP425), (2) broad band FITC or GFP (Chroma I3: BP450-490/510/LP515, (3) narrow band FITC or GFP (Chroma +L5: 480/40/505/BP527/30, (4) rhodamine (Chroma N2.1: BP515-560/580/LP590).
Light microscopy auxiliary equipment includes two microtomes for parafin sectioning and a Leica TP1020 tissue processor for sample preparation.
Instrumentation. The center has a Leica TCS SP confocal scanning microscope. in this "SP" model a specrophotometer replaces emission filters giving ultimate flexibility for "filter-design". This feature is particularly advantegeous for difficult dye combinations ,or plant tissue, where autofluorescence may be a problem. This microscope has three excitation lasers that cover excitation ranges of the most commonly used fluorescent dyes: (1) Argon (450nm and 488nm), (2) Krypton (568 nm), and (3) Helium-Neon (630 nm).
The facility staff provides individual training sessions for confocal microscope users. Users must be trained by the MCIC personnel in order to use the confocal microscope.
Transmission Electron Microscopy
Instrumentation. Two transmission electron microscopes are maintained at the center. A Hitachi H-7500 is characterized by a new electron optical lens system, which allows the acquisition of high contrast images even at low resolution. It operates in a Windows environment and allows user friendly interactive operation. Its magnifications ranges from 700X to 600.000X, with accelerating voltage between 40kV and 120kV.
The Philips 201 is suitable for routine biological microscopy, and it offers moderately high-resolution for standard TEM viewing of biological samples. Accelerating voltage ranges between 40kV and 100kV, and magnification between 500X and 100,000X.
Sample preparation techniques routinely available at the center:
Ultra-thin sectioning for morphology study. For ultrastuctural and morphological studies, the biological specimen is fixed, dehydrated and embedded in plastic blocks, which are then sectioned into app. 80 NM sections. After staining with electron dense stained, the sections are observed at the electron microscope. We use a generic fixation and embedding protocol which produces a good fixation and ultra-structural preservation. If you would like to use a different protocol, you need to talk to us.
Negative staining. This is a staining technique that allows visualization of particulate biological samples in suspension, such as virus particles, bacteria, bacteriophages and protein or lipoprotein complexes. An electron dense stain containing heavy ions, such as uranyl acetate or lead citrate, is applied to the to the biological sample in suspension. The stain deposits around the specimen, which is then placed onto formovar carbon coated grids for electron microscope observation.
Immunogold labeling. This is a very powerful technique that provides information on the identity, localization and distribution of macromolecules at an ultrastructural level. The techniques involved in this procedure vary and no single preparative procedure can be generally applied and the best method has to be worked out for each experimental system.
Instrumentation. Our Hitachi S-3500N scanning electron microscope can operate in both conventional high vacuum in the specimen chamber, or at elevated pressure. This feature allows for minimal specimen preparation and allows viewing of hydrated biological samples without the conductive coatings required for conventional scanning electron microscopy. A resolution of 3.0nm is obtainable in high vacuum mode, and a resolution of 4.5nm in variable-pressure (VP) mode. The microscope is PC controlled and allows quality digital image recording and storage. The microscope is also equipped with a Noran X-ray analytical device for dispersive X-ray elemental microanalysis.
MCIC users have also access to the USDA cold field emission Hitachi S-4700 scanning electron microscope, which is scanning electron microscope. This microscope is also equipped with a Noran X-ray dispersive analysis device. If you would like to use this microscope, you need to contact:
Charles R. Krause, Ph.D., Adjunct Associate Professor, USDA-ARS, krause.2@osu.edu, or
Leona Horst, Technical Assistant, USDA-ARS, horst.9@osu.edu
Services. We provide a variety of sample preparation techniques and help users with critical point drying and sample coating with carbon, gold or platinum.
Click on the images to access the gallery
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Viruses |
Spiroplasma |
Bacteria |
Protozoa |
Fungi & Oomyces |
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Insects |
Higher plants |
Plant Cells |
Animal Cells |
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